<p><b>AIM</b>To develop a
method for assessing
sperm function by measuring released
acrosin activity during the
acrosome reaction (AR).</p><p><b>
METHODS</b>
Human semen samples were obtained from 24 healthy
donors with proven
fertility after 3-7 days of
sexual abstinence. After collection, samples were liquefied for 30 min at room
temperature. Standard
semen parameters were evaluated according to
World Health Organization (
WHO) criteria.
Calcium ionophore A23187 and
progesterone (P4) were used to stimulate the
sperm to undergo AR.
After treatment,
sperm were incubated with the supravital
dye Hoechst33258, fixed in a
glutaraldehyde-
phosphate-buffered
saline solution, and the acrosomal status was determined by
fluorescence microscopy with
fluorescein isothiocyanate-labeled
Pisum sativum agglutinin (
FITC-PSA). The percentage of
sperm undergoing AR (AR%) was compared to
sperm acrosin activities as assessed by spectrocolorimetry. The correlation between AR% and
acrosin activity was determined by
statistical analysis.</p><p><b>RESULTS</b>The AR% and released
acrosin activity were both markedly increased with
A23187 and P4 stimulation.
Sperm motility and viability were significantly higher after stimulation with P4 versus stimulation with
A23187 (P < 0.001). There was a significant positive correlation between released
acrosin activity and AR% determined by
FITC-PSA
staining (r=0.916, P < 0.001).</p><p><b>CONCLUSION</b>Spectrocolorimetric measurement of released
acrosin activity might serve as a reasonable alternative
method to evaluate AR.</p>